Research On The Cardioprotective Effect Of Guanxin ⅡSubstance Basis And Its Vasodilative Effect And Mechanism In Thoracic Aorta Of Rats

BACKGROUND:The main form of Traditional Chinese Medicine (TCM) is composite prescription. Its therapeutic effects are confirmed after over thousand years clinical practice. It remains largely a secret that which and how active constituents of composite prescription produce the effectiveness, which has now become a main bottleneck restricting the TCM modernizations. We will study the cardioprotective effect of Guanxin II substance basis and its vasodilative effect and mechanism on thoracic aorta of rats with bioethnopharmaceutical analytical phannacology and solve the problem which has puzzled TCM for so long time。I Review the research on the cardioprotective effect of Guanxin II substance basisObjective To review the cardioprotective effects of GXII and FATPE, alone and in combination, and of some components of FATPE in AMI rat. Method Using my task group material, SD rats with acute myocardial infarction (AMI) were induced by coronary occlusion.The contents of the five components including ferulic acid (0.169±0.004), tanshinol (0.936±0.013), hydroxyl safflor yellow A (2.591±0.021), protocatechualdehyde (0.018±0.001) and Paeoniflorin (3.715±0.123) in the formula were determined by high-performance liquid chromatography (HPLC) method respectively. Compare the cardioprotect effects of GXII and FATPE, alone and in combination, and of some components of FATPE in AMI rat. Result Comparing the mixture of FTA and original formula, FAT(dose=amount of the original formula) was similar to GXII in cardioprotective effects. Conclusion The cardioprotective effect substance basis of Guanxin II is FTA.II Reasearch on vasodilative effect and mechanism of Tanshinol in thoracic aorta of ratsObjective The aim was to study Tanshinol(10-1M～10-6M) in isolated rat thoracic aortic ring vasodilative effect and its mechanisms. Method Isometric tension was recorded in response to drugs in organ bath. Effects of Tanshinol(10-1M～10-6M) on the vascular tone of resting, high KCl(60mmol/L) and PE(1μmol/L) pre-constricted rat thoracic aorta with or without endothelium were determined. To explore the mechanism, nitric oxide synthase inhibitor L-NAME(10μmol/L), guanylyl cyclase inhibitor methyline blue MB(10μmol/L), cyclooxygenase inhibitor indomethacin(10μmol/L) were ued. Result No vasomotor response to accumulated Tanshinol(10-1M～10-6M) were recorded in both resting and KCl-depolarized rings Tanshinol(10-1M～10-6M) caused concentration relaxation of aorta rings preconstricted with PE(1μmol/L) in endothelium-intact rings. Removal of the endothelium, or pretreated with L-NAME(10μmol/L) or MB(10μmol/L) inhibited the relaxation of Tanshinol(10-1M～10-6M). Pretreatment with Indo (10μmol/L) could not abolish the relaxation response to Tanshinol. Conclusion The results indicated that the relaxation by Tanshinol(10-1M～10-6M) in rat aorta ring is endothelium dependent and is possibly mediated by the NO-sGC-cGMP pathway.Ⅲ Reasearch on vasodilative effect and mechanism of Ferulic acid in thoracic aorta of ratsObjective The aim was to study Ferulic acid(10-1mM～103mM) in isolated rat thoracic aortic ring vasodilative effect and its mechanisms. Method Isometric tension was recorded in response to drugs in organ bath. Effects of Ferulic acid(10-1mM～103mM) on the vascular tone of resting, high KCl(60mmol/L) and PE(1μmol/L) pre-constricted rat thoracic aorta with or without endothelium were determined. In some experiments, Effects of Ferulic acid(10-1mM～103mM) on the vascular tone of PE(1μmol/L)and Caf(20mol/L) pre-constricted rat thoracic aorta in Ca2+-free K-H solution were also determined. To explore the further mechanism on K+channels, ATP-Sensitive potassium channel inhibitor Gli(10μmol/L), inward rectifier potassium channel BaCl2(lmmol/L), calcium activated potassium channel TEA(10mmol/L), voltage dependent potassium channel4-AP(1mmol/L) were also used. Result No vasomotor response to accumulated Ferulic acid(10-1mM～103mM) were recorded in both resting and KCl-depolarized rings. Ferulic acid (10-1mM～103mM) caused concentration relaxation of aorta rings preconstricted with PE(1μmol/L) in endothelium-intact rings. Removal of the endothelium, or pretreated with L-NAME(10μmol/L) did not inhibite the relaxation of Ferulic acid(10-1mM～103mM). Pretreatment with Indo(10μmol/L) could not abolish the relaxation response to Ferulic acid(10-1mM～103mM). In Ca2+-free K-H solution, Ferulic acid(10-1mM～103mM) could relax Caf-constricted rat thoracic aorta. Gli(10μmol/L) could partly abolish the relaxation response to Ferulic acid. Conclusion The results indicated that the relaxation by Ferulic acid(10-1mM～103mM) in rat aorta ring is endothelium independent and is possibly mediated by blocking ROC and furtherly inhibiting ryanodine or caffeine sensitive calcium release, partly open ATP-Sensitive potassium channel.IV Reasearch on vasodilative effect and mechanism of hydroxyl safflor yellow A in thoracic aorta of ratsObjective The aim was to study hydroxyl safflor yellow A(10-6M～10-1M) in isolated rat thoracic aortic ring vasodilative effect and its mechanisms. Method Isometric tension was recorded in response to drugs in organ bath. Effects of hydroxyl safflor yellow A on the vascular tone of resting, high KCl(60mmol/L) and PE(1μmol/L) pre-constricted rat thoracic aorta with or without endothelium were determined. To explore the mechanism, nitric oxide synthase inhibitor L-NAME and cyclooxygenase inhibitor indomethacin were ued. In some experiments, Effects of hydroxyl safflor yellow A(10-6M～10-1M) on the vascular tone of PE(1μmol/L)and Caf(20mol/L) pre-constricted rat thoracic aorta in Ca2+-free K-H solution were also determined. To explore the further mechanism on K+channels, ATP-Sensitive potassium channel inhibitor Gli(10μmol/L), inward rectifier potassium channel BaCl2(lmmol/L), calcium activated potassium channel TEA(1Ommol/L), voltage dependent potassium channel4-AP(1mmol/L) were also used. Result No vasomotor response to accumulated hydroxyl safflor yellow A were recorded in both resting and KCl-depolarized rings. Hydroxyl safflor yellow A(10-6M～10-1M) caused concentration relaxation of aorta rings preconstricted with PE(1μmol/L) in endothelium-intact rings. Removal of the endothelium, or pretreated with Indo could partly inhibite the relaxation of hydroxyl safflor yellow A(10-6M～10-1M). Pretreatment with L-NAME(10μmol/L) could not abolish the relaxation response to hydroxyl safflor yellow A(10-6M～10-1M). In Ca2+-free K-H solution, hydroxyl safflor yellow A(106M～10-1M) could relax PE-constricted rat thoracic aorta.4-AP could partly abolish the relaxation response to hydroxyl safflor yellow A(10-6M～10-1M). Conclusion In conclusion, hydroxyl safflor yellow A(106M～10-1M) relaxed rat aorta rings through blocking ROC, VOC and furtherly inhibiting IP3sensitive calcium release, partly open voltage dependent potassium channel. The endothelium-dependent vessel dilation of hydroxyl safflor yellow A(10-6M～10-1M) was attributed mainly to the endothelium-dependent cyclooxygenase pathway. Conclusion GXII is multi-target therapy in cardioprotective effect, and FTA was proved to be the pharmacodynamics substance basis. FAT was similar to GXII in increasing MBF. The combination mechanisms of FTA is the GX II vasodilative mechanism. GXII relaxed thoracic aorta of rats through blocking calcium influx and intracellular calcium release extracellularly, inhibiting Ca2+release in IP3, ryanodine or Caffeine sensitive calcium pool intracellularly and mediating by opening of ATP-dependent potassium channels and voltage dependent potassium channel. The endothelium-dependent vessel dilation of GXII was attributed mainly to the endothelium-dependent NO-sGC-cGMP and cyclooxygenase pathway.